Titanium dioxide nanoparticle coating of polymethacrylate-based chromatographic monoliths for phosphopetides enrichment

Metal oxide affinity chromatography has been one of the approaches for specific enrichment of phosphopeptides from complex samples, based on specific phosphopeptide adsorption forming bidentate chelates between phosphate anions and the surface of a metal oxide, such as TiO₂, ZrO₂, Fe₂O₃, and Al₂O₃. Due to convective mass transfer, flow-independent resolution and high dynamic binding capacity, monolith chromatographic supports have become important in studies where high resolution and selectivity are required. Here, we report the first synthesis and characterization of immobilisation of rutile TiO₂ nanoparticles onto organic monolithic chromatographic support (CIM-OH-TiO₂). We demonstrate the specificity of CIM-OH-TiO₂ column for enrichment of phosphopeptides by studying chromatographic separation of model phosphorylated and nonphosphorylated peptides as well as proving the phosphopeptide enrichment of digested bovine α-casein. The work described here opens the possibility for a faster, more selective enrichment of phosphopeptides from biological samples that will enable future advances in studying protein phosphorylation.     

膜富集-紫外光谱法快速检测自来水中痕量2-萘酚

基于紫外光谱结合膜富集技术,提出了一种快速、灵敏检测自来水中2-萘酚的方法。以尼龙滤膜(孔径0.22μm)为富集材料,将富集到膜上的2-萘酚不经洗脱直接测定其漫反射紫外光谱,以实现定量分析目的。该方法方便快捷,大大提高了灵敏度。对p H值、样品体积和真空度等条件进行优化,并对方法的分析性能进行了评价。结果显示,2-萘酚的浓度与吸光度(波长为331 nm)在0.3~10 mg/L范围内用二次函数可得到良好的拟合关系,在0.3~1.0 mg/L和1.0~10 mg/L范围内呈分段的线性关系,相关系数(r2)分别为0.999 1和0.995 5。在最佳条件下,检出限达0.265 6 mg/L。该方法准确度和精密度较理想,适用于对自来水中2-萘酚的检测。     

基于液相色谱-串联质谱技术的磷酸化蛋白质组学分析

本文运用稳定同位素双甲基化标记技术、固定相金属离子螯合层析(IMAC)技术并结合液相色谱-串联质谱法研究了仙台病毒感染引起的宿主细胞磷酸化蛋白质组变化。实验发现定量的4 289个磷酸化肽段有~20%的蛋白质磷酸化位点发生了显著变化;通路富集分析表明哺乳动物雷帕毒素靶蛋白(mTOR)通路和剪接体(Spliceosome)通路可能参与宿主细胞对病毒的应答;通过对显著变化磷酸化肽段进行基序分析,发现了9个代表性的磷酸化修饰位点基序。本研究方法对于深入解析抗病毒天然免疫信号通路提供了新线索。     

Experimental choices for the determination of carbonyl compounds in air

The analysis of carbonyls in ambient air has received a great deal of scientific attention with the advancement of analytical techniques and increased demand for the build-up of its data base. In this review article, we have attempted to provide some insight into the relative performance of different instrumental approaches available for the analysis of ambient carbonyls with a major emphasis on high performance liquid chromatographic and gas chromatographic methods. Reported in several international standard procedures, derivatization of carbonyls with 2,4-dinitrophenylhydrazine (2,4-DNPH) with either an impinger or cartridges is the most commonly used method of HPLC detection. In this respect, a number of alternative hydrazine reagents have also been discussed for use with HPLC. In contrast, GC methods based on the combined application of adsorptive enrichment on solid sorbents and thermal desorption are examined with regard to their suitability for carbonyl analysis in air. Particular emphasis has been directed towards the advantages and drawbacks of these different instrumental techniques for ambient carbonyls. Based on this comparative approach, we discuss the suitability for each method for carbonyl analysis.     

Identification of phosphoproteins and determination of phosphorylation sites by zirconium dioxide enrichment and SELDI-MS/MS

Reversible protein phosphorylation mediated by protein kinases and phosphatases is the most studied post-translational modification. Efficient characterization of phosphoproteomes is hampered by (1) low stoechiometry, (2) the dynamic nature of the phosphorylation process and (3) the difficulties of mass spectrometry to identify phosphoproteins from complex mixtures and to determine their sites of phosphorylation. Combination of the phosphopeptide enrichment method with MALDI-TOFMS, or alternatively, with HPLC-ESI-MS/MS and MS(3) analysis was shown to be a step forward for the successful application of MS in the study of protein phosphorylation. In our study we used phosphopeptide enrichment performed in a simple single-tube experiment using zirconium dioxide (ZrO(2)). A simple protein mixture containing precipitated bovine milk caseins was enzymatically digested and the mixture of tryptic fragments was analysed before and after enrichment using nanoflow HPLC-ESI-MS/MS and surface-enhanced laser desorption/ionization (SELDI)-MS/MS on QqTOF instruments to compare the efficiency of the two methods in the determination of phosphorylation sites. Both approaches confirm the high selectivity obtained by the use of batch-wise, ZrO(2)-based protocol using di-ammonium phosphate as the eluting buffer. More phosphorylation sites (five for beta-casein and three for alpha(S1)-casein) were characterized by SELDI-MS/MS than by nanoflow HPLC-ESI-MS/MS. Therefore, ZrO(2)-based phosphopeptide enrichment combined with SELDI-MS/MS is an attractive alternative to previously reported approaches for the study of protein phosphorylation in mixtures of low complexity with the advance of fast in situ peptide purification. The method was limited to successful analysis of high-abundance proteins. Only one phosphorylation site was determined for the minor casein component alpha(S2)-casein by ESI-MS/MS and none for kappa-casein. Therefore an improvement in enrichment efficiency, especially for successful phosphoproteomic applications, is needed.     

壳聚糖富氧膜的研究(Ⅰ)

壳聚糖可用来进行醇水分离,但用于氧氮分离尚未见报道.本文成功地以壳聚糖复合膜分离空气中的氧氮,并且发现干燥膜与在干燥膜表面涂上一层水的“湿膜”之间存在明显的氧氮分离差别. 1实验部分壳聚糖(浙江玉环化工厂)经水洗、酸溶、重沉淀处理后,用稀醋酸溶解,配成一定浓度的     

用于昆虫外激素研究的微量顶空收集及热解吸气相色谱进样方法初探

描述了一个结合项空气流收集与无溶剂热解吸气相色谱进样的方法。用填充PorapskQ的微量注射器作为吸附管进行气流收集。将收集物不经溶剂洗脱直接进行热解吸进样。用人工合成的昆虫外激素化合物反-7-十二碳单烯乙酸酯（E-7-DA）及顺-5,反-7-十二碳二烯乙酸酯（Z-5,E-7-DDA）测定了方法的回收率,初步探索了运用于昆虫外激素分析的可行性,并讨论了提高回收率的途径。     

气相色谱分析样品的提取与富集技术

对几种常用或新近发展起来的气相色谱分析样品的提取和富集方法如固相萃取法、吹扫——捕集法、热解析法等作了简要论述,着重讨论了它们的应用特点、优缺点,并对这几种提取、富集方法进行了比较。     

泡沫塑料富集-火焰原子吸收光谱法测定金矿石中金

采用聚氨酯泡塑富集硫脲洗脱-火焰原子吸收光谱法测定金矿石中金。对样品称样量、硫脲洗脱时间、磷酸三丁酯的吸附效果和铁元素的干扰等工作条件进行了优化,解决了泡沫塑料吸附效率低和铁元素的干扰等技术性难题,提高了分析结果的精密度和准确度,弥补了传统的泡沫塑料富集-火焰原子吸收光谱法测定金矿石中金量的不足,方法检出限为0.06 μg/g,测定范围为 0.2 μg/g～100 μg/g,测定结果与标准值符合性较好,无显著性差异,精密度(RSD, n = 9) 小于2%。方法具有快速、简便、实用等优点,分析误差满足常规化学分析法的要求,能满足金矿石中金量的快速、准确测定。     

成都平原典型地区水果-土壤重金属污染特征研究

为摸清成都平原土壤-水果系统中重金属污染水平和富集特征,通过对成都平原典型地区333个水果作物及其根部表层土壤中Hg、As、Pb、Cd、Cu、Cr和土壤理化指标的测定,结果表明：（1）研究区土壤养分整体呈现出有效磷和碱解氮较丰富,有机质和速效钾较缺乏的特征,可通过增施有机肥、地面覆草、施用绿肥等方式提升土壤肥力水平。（2）研究区土壤重金属存在部分点位超标,以Cd的超标率最高。土壤重金属单因子评价污染等级为清洁,土壤综合污染指数评价污染等级为中度污染,其主要贡献为Cd。研究区重金属存在一定程度的富集,Cd和Hg的污染指数高,潜在生态风险强,应重点关注土壤Cd污染和Hg含量水平的增加趋势,将Cd污染作为下一步土壤风险管控和修复治理的重点目标。（3）研究区水果Pb、Cd的超标率分别为5.7%、7.8%,不同水果Pb、Cd的超标率差异显著,草莓Cd的超标率最高。研究区水果中重金属未对成人构成非致癌健康风险,而水果中的As和Cu对儿童构成了非致癌健康风险,As和Cu对水果重金属非致癌总指数的贡献超过70%,应特别关注水果中As和Cu的管控,以免造成较大的食源性危害。（4）重金属在水果与土壤中的含量水平关联性较低,水果中重金属的含量与水果富集重金属的能力关联度极高,应进一步查明水果重金属与土壤重金属形态的关系,并结合调整水果种植结构、筛选重金属低积累水果品种等方式降低水果重金属的健康风险。